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Determination of inosine 5'-monophosphate dehydrogenase activity by high performance liquid chromatography in comparison with normalization methods |
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| รหัสดีโอไอ | |
| Creator | 1. MT. Nguyen 2. NT. Tran 3. M. Vincent 4. A. Capron 5. M. Mourad 6. P. Wallemacq |
| Title | Determination of inosine 5'-monophosphate dehydrogenase activity by high performance liquid chromatography in comparison with normalization methods |
| Publisher | Faculty of Pharmacy, Mahidol University |
| Publication Year | 2558 |
| Journal Title | Mahidol University Journal of Pharmaceutical Sciences |
| Journal Vol. | 42 |
| Journal No. | 4 |
| Page no. | 162-168 |
| Keyword | inosine 5'-monophosphate dehydrogenase activity,mycophenolic acid,HPLC,peripheral blood mononuclear cells |
| Abstract | Mycophenolic acid (MPA) is a potent, selective and uncompetitive inhibitor of the inosine 5'-monophophate dehydrogenase (IMPDH), which is the rate-limiting enzyme in the de novo synthesis of guanosine nucleotides being required during T and B lymphocyte proliferation. In this study, HPLC method was developed and validated to determine the IMPDH activity in human peripheral blood mononuclear cells (PBMCs) for the purpose of investigating the effect of MPAon lymphocytes. PBMCs were isolated from total blood samples. IMPDH activity in PBMCs was expressed according to two normalisation methods, namely as the ratio of produced xanthosine monophosphate (XMP) (micromoles) versus either adenosine monophosphate (AMP) (moles), or protein (mg) concentration per incubation time. IMPDH activities were assessed in 41 healthy volunteers. The total HPLC run time was 14 min. This method was linear in the range of 0.5-80 ?mol/L for both AMP and XMP. |
